Methods for producing Ex vivo models for inflammatory disease and uses thereof

ABSTRACT

The invention relates to methods for inducing a pro-asthma/pro-inflammatory like state in a resident tissue cell, comprising contacting the cell with one or more cytokines, e.g., IL-1β, TNFα or both. Methods are also disclosed for identifying genes that regulate responses to anti-inflammatory drugs, to methods for drug screening, and to methods for identifying genes that correlate with various pro-asthma/pro-inflammatory disease phenotypes.

RELATED APPLICATION

This application is a divisional of U.S. application Ser. No.09/947,954, filed Sep. 6, 2001.

The entire teachings of the above application are incorporated herein byreference.

GOVERNMENT SUPPORT

The invention was supported by grants HL-59906, HL-31467, HL-58245 andHL-61038 from the National Heart, Lung and Blood Institute. TheGovernment has certain rights in the invention.

BACKGROUND OF THE INVENTION

The pleiotropic cytokines, IL-1β and TNFα, among other effectors, havebeen implicated in the pathophysiology of asthma and other inflammatorydiseases. Altered airway responsiveness to bronchoactive constrictor andrelaxant stimuli is the characteristic pathophysiological feature ofbronchial asthma. While infiltration of the airways with inflammatorycells, principally involving eosinophils, mast cells, and lymphocytes ischaracteristic of altered airway responsiveness, airway smooth muscle(ASM) itself has the capacity to autologously induce changes in itsconstrictor and relaxant responsiveness secondary to the induced releaseand autocrine actions of certain pro-inflammatory cytokines. Forexample, IgE-dependent atopic sensitization and rhinovirus inoculationof ASM provoke the release of Th1 and Th2-type cytokines, IL-1β, andother cytokines from the ASM itself; and these cytokines acting alone orin combination elicit changes in ASM responsiveness.

Effectors such as cytokines typically are involved in a broad class ofsignaling events. Indeed, altered levels in IL-1β, and TNFα signalingactivity are observed in inflammatory diseases other than asthma aswell.

SUMMARY OF THE INVENTION

The present invention relates to methods for determining a patient'sresponsiveness to treatment for asthma or related inflammatoryconditions.

In one embodiment, the invention is directed to a method for inducing apro-inflammatory like state in a resident tissue cell, comprisingcontacting the cell with a cytokine that induces a pro-inflammatory likestate such as, for example, IL-1β, TNFα or both. Cells induced toexhibit a pro-inflammatory like state can be, for example, airway smoothmuscle cell, epithelial cell, keratinocyte, synovial cell, glial celland villous intestinal cell. The pro-inflammatory like state can be apro-asthma like state.

In another embodiment, the invention is directed to a resident tissuecell induced to exhibit a pro-inflammatory like state according to themethods described herein. Resident tissue cells can be, for example,airway smooth muscle or airway epithelial cells.

In another embodiment, the invention is directed to a method forscreening drug candidates for treating an inflammatory disease,including: contacting a resident cell induced to exhibit apro-inflammatory like state according to the methods described hereinwith a drug candidate for treating the inflammatory disease; andassaying for a pro-inflammatory like state, such that an absence of thepro-inflammatory like state is indicative of the drug candidate beingeffective in treating the inflammatory disease. In a particularembodiment, the inflammatory disease can be asthma, atopy, rheumatoidarthritis, psoriasis, inflammatory bowel disease (IBD) and chronicobstructive pulmonary disease (COPD). Atopy can be rhinitis,conjunctivitis, dermatitis and eczema.

In another embodiment, the invention is directed to a method forscreening drug candidates for treating an inflammatory disease,including: contacting a resident tissue cell induced to exhibit apro-inflammatory like state according to the methods described hereinwith a drug candidate for treating an inflammatory disease; and assayingfor a pro-inflammatory like state, such that an absence of thepro-inflammatory like state is indicative of the drug candidate beingeffective in treating an inflammatory disease. In a particularembodiment, the inflammatory disease can be asthma, atopy, rheumatoidarthritis, psoriasis, IBD and COPD.

In another embodiment, the invention is directed to a method ofidentifying genes associated with an inflammatory disease, including:obtaining resident tissue cells induced to mimic the inflammatorydisease; assaying the expression level of at least one gene in thecells; comparing the expression level to the baseline expression levelsin cells not induced to mimic the inflammatory disease; and identifyinga difference in expression level in cells induced to mimic theinflammatory disease versus cells that do not mimic the inflammatorydisease, such difference indicating the gene is associated with theinflammatory disease. In another embodiment, the invention is directedto a method for identifying genes that are involved in regulating drugresponses and present candidate genes for development of new therapy fortreating an inflammatory disease, including: contacting a cell inducedto exhibit a pro-inflammatory like state according to the methodsdescribed herein with a drug candidate for treating the inflammatorydisease; and assaying for a pro-inflammatory like state, such that geneswhose expression correlates with an absence of the pro-inflammatory likestate are indicative of the gene being involved with regulating theresponse to the drug in treating the inflammatory disease. In oneembodiment, the inflammatory disease can be asthma, atopy (e.g.,rhinitis, conjunctivitis, dermatitis or eczema), rheumatoid arthritis,psoriasis, IBD or COPD. In one embodiment, the informative gene can beselected from the genes described in Tables 1 and 2.

In yet another embodiment, the invention is directed to a method fordiagnosing an inflammatory disease, including: obtaining or generating agene expression profile from a sample for at least one informative geneidentified by methods described herein; comparing the expression profileof the informative gene to a reference expression profile for theinformative gene in a cell induced for pro-asthma/pro-inflammatory likeconditions; and comparing the expression profile of the informative geneto a reference expression profile for the informative gene in a cellthat does not exhibit pro-asthma/pro-inflammatory like conditions,wherein similarity between the sample expression profile of theinformative gene and either of the reference expression profiles allowsfor a positive or negative diagnosis of the patient from whom the samplewas obtained. In one embodiment, the informative gene can be selectedfrom the genes described in Tables 1 and 2.

In yet another embodiment, the invention is directed to an expressionprofile indicative of the presence of asthma in a patient, including atleast one informative gene of Table 1 and Table 2.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing and other objects, features and advantages of theinvention will be apparent from the following more particulardescription of preferred embodiments of the invention, as illustrated inthe accompanying drawings.

FIG. 1 is a graphical comparison of ASM constrictor responses toacetylcholine (ACh) in control (open symbols) and IL-1β/TNFα treated(filled symbols) ASM tissues. Data represent means±SE from six pairedexperiments. Relative to tissues incubated with media alone, bothT_(max) and ED₅₀ responses to ACh were significantly enhanced (p<0.01and <0.05, respectively) in ASM tissues that were co-incubated withIL-1β/TNFα, combined (filled symbols).

FIG. 2 is a graphical comparison of airway relaxant responses toisoproterenol in control (open symbols) and IL-1β/TNFα treated (filledsymbols) ASM tissues. Data represent means±SE from six pairedexperiments. Relative to tissues incubated with media alone, bothR_(max) and pD₅₀ responses to isoproterenol were significantlyattenuated (p<0.01 and <0.05, respectively) in ASM tissues that weretreated with IL-1β/TNFα, combined (filled symbols).

FIG. 3 is a graph showing ASM mRNA expression of 25 cytokine/chemokinegenes demonstrating >2-fold change in expression following 4 hr exposureto IL-1β/TNFα combined, using gene array technology representingapproximately 5000 genes. Each gene is identified by its gene symbol andGenBank accession number, and plotted in relation to its respectivemagnitude (mean±SE values) of fold-change in expression from baselinevalues.

FIG. 4 is a graph showing ASM mRNA expression of 8 celladhesion/extracellular matrix genes demonstrating >2-fold change inexpression following 4 hr exposure to IL-1β/TNFα combined, using theHu95GeneFL array from Affymetrix. Each gene is identified by its genesymbol and GenBank accession number, and plotted in relation to itsrespective magnitude (mean±SE values) of fold-change in expression frombaseline values.

FIG. 5 is a graph showing ASM mRNA expression of 14 transcription factorgenes demonstrating >2-fold change in expression following 4 hr exposureto IL-1β/TNFα combined, using the Hu95GeneFL array. Each gene isidentified by its gene symbol and GenBank accession number, and plottedin relation to its respective magnitude (mean±SE values) of fold-changein expression from baseline values.

FIG. 6 is a graph showing ASM mRNA expression of 18 cellsignaling/metabolism-related genes demonstrating >2-fold change inexpression following 4 hr exposure to IL-1α a combined, using theHu95GeneFL array. Each gene is identified by its gene symbol and GenBankaccession number, and plotted in relation to its respective magnitude(mean±SE values) of fold-change in expression from baseline values.

FIG. 7 is a graphical comparison of ASM constrictor responses to ACh incontrol (open circles) and IL-1β/TNFα treated ASM tissues in the absence(filled circles) and presence (filled squares) of pretreatment withdexamethasone 10⁻⁵M. Data represent means±SE from six pairedexperiments. Relative to tissues incubated with media alone, both theT_(max) and ED₅₀ responses to ACh were significantly enhanced in ASMsegments that were exposed to IL-1β/TNFα, whereas the latter effects onthe T_(max) and ED₅₀ values were largely prevented by pre-treating theASM tissues with dexamethasone (p≦0.01 and p≦0.05, respectively). Incontrast, treatment with dexamethasone 10⁻⁵M alone (open squares), hadno effects on either the T_(max) or ED₅₀ responses to ACh.

FIG. 8 is a graphical comparison of ASM relaxant responses toisoproterenol in control (open circles) and IL-1β/TNFα treated ASMtissues in the absence (filled circles) and presence (filled squares) ofpretreatment with dexamethasone 10⁻⁵M. Data represent means±SE from sixpaired experiments. Relative to tissues incubated with media alone, boththe R_(max) and pD₅₀ responses to isoproterenol were significantlyenhanced in ASM segments that were exposed to IL-1β/TNFα, whereas thelatter effects on the R_(max) and pD₅₀ values were largely prevented bypre-treating the ASM tissues with dexamethasone (p≦0.01 and p≦0.05,respectively). In contrast, treatment with dexamethasone 10⁻⁵M alone(open squares), had no effects on either the R_(max) or pD₅₀ responsesto isoproterenol.

FIG. 9 is a graph showing the effects of dexamethasone on IL-1β/TNFαinduced gene expression in human ASM cells, using a human DNA gene chipfrom Affymetrix. All genes belonging to the four categories of genesshown in FIGS. 3-6 are illustrated. Data representdexamethasone-mediated mean inhibition (MER=1.0) and mean enhancement(MER>1.0) of mRNA expression from maximum levels induced by IL-1β/TNFαtreatment from 2 separate experiments.

DETAILED DESCRIPTION OF THE INVENTION

A description of preferred embodiments of the invention follows.

The present invention is directed to methods for inducing cells toexhibit pro-inflammatory like characteristics, to methods for drugscreening, to methods for identifying genes that are differentiallyexpressed in cells displaying pro-inflammatory like characteristicsversus normal cells, and to the genes thus identified. The invention isbased upon the discovery that cultured cells can be induced to mimic apro-inflammatory like state. The methods disclosed herein, in part,refer to the activation of the IL-1β/TNFα signaling pathway, theactivation of which is shared among inflammatory diseases in addition toasthma. Hence, as used herein, “pro-asthma” like characteristics referto “pro-inflammatory” like characteristics in particular cells affectedby asthma. As used herein, “pro-inflammatory like” is a description of astate or characteristics associated with inflammatory diseases ingeneral with asthma being a well characterized and well studied exampleof inflammatory diseases. Thus, pro-asthma like characteristics are alsopro-inflammatory like characteristics and, where referred to herein, aremeant to describe characteristics caused by differential levels ofeffectors specific to inflammatory disease. The term is not limiting toconditions associated specifically with asthma. However, as asthma is awell characterized example of inflammatory disease, characteristicsassociated with asthma are also common for other inflammatory diseases.

Asthma, or Reversible Obstructive Airway Disease (ROAD), is a conditionin which the airways of the lungs become either narrowed or completelyblocked, impeding normal breathing and leading to potentially moresevere health problems. Although normal airways have the potential forconstricting in response to allergens or irritants, the asthmatic'sairways are oversensitive or hyper-reactive. In response to stimuli, theairways may become obstructed by one of the following: constriction ofthe muscles surrounding the airway; inflammation and swelling of theairway; or increased mucus production which clogs the airway. Once theairways have become obstructed, it takes more effort to force airthrough them, so that breathing becomes labored. Because exhalingthrough the obstructed airways is difficult, too much stale air remainsin the lungs after each breath. This accumulation of stale air decreasesthe amount of fresh air that can be taken in with each new breath, sonot only is there less oxygen available for the whole body, the highconcentration of carbon dioxide in the lungs causes the blood supply tobecome acidic as well. This acidity in the blood may rise to toxiclevels if the asthma remains untreated.

Although asthma creates difficulties in breathing and can lead to moreserious problems, the lung obstruction associated with asthma isreversible, either spontaneously or with medication. Asthmatics can takeanti-inflammatory agents such as corticosteroids, brochodilators andleukotriene antagonists reduce inflammation and asthma symptoms.Although asthma has been treated by these methods for several years, asignificant fraction of asthma patients are resistant to treatment. Asthere are risks associated with methods for treating asthma,identification of patients that will be responsive to treatment isimportant. Methods described herein are used to identify genes thatregulate drug response. Described herein are methods for inducingconditions in cultured cells that mimic asthma and inflammatory diseaseconditions and methods for utilizing said cultured cells in methods fordiagnosing disease, drug screening and obtaining expression profiles.

Corticosteroids are sometimes also referred to as “steroids.” This typeof medication is not related to the anabolic steroids that are misusedby some athletes to increase performance. Rather, corticosteroids havebeen used as a treatment for asthma and allergies since 1948. Theydecrease airway inflammation and swelling in the bronchial tubes; reducemucus production by the cells lining the bronchial tubes; decrease thechain of overreaction (hyper-reactivity) in the airways; and assist theairway smooth muscle to respond to other medications such asbeta-agonist drugs. Corticosteroids can be administered in a variety ofways, such as through the use of an inhaler, topically, orally, orthrough injection. Topical preparations (on specific surface areas suchas skin or the lining of the bronchial tubes) may be applied as creamsor sprays (inhalers). Corticosteroid inhalers are recommended forpatients with daily, moderate or severe asthma symptoms. Oralcorticosteroids and injected corticosteroids are generally onlyprescribed for those with severe asthma symptoms.

Although the use of corticosteroids has been commonplace for severalyears, they are not always effective and significant side effects dooccur. Some people experience minor side effects of hoarseness andthrush (a fungal infection of the mouth and throat) from usingcorticosteroid inhalers. Also, long-term use of inhaled corticosteroidshas been implicated in reduced growth velocity in children. Oralcorticosteroids can have more side effects than inhaled corticosteroids.Oral corticosteroids are prescribed for long durations only when othertreatments have failed to restore normal lung function and the risks ofuncontrolled asthma are greater than the side effects of the steroids.For example, prednisone, one of the most commonly prescribedcorticosteroids, can lead to possible side effects of weight gain,increased appetite, menstrual irregularities and cramps, heartburn, andindigestion. Some patients experience side effects such as loss ofenergy, poor appetite, and severe muscle aches or joint pains when theirdosage of cortisone tablets is decreased. Long-term oral corticosteroiduse may cause side effects such as ulcers, weight gain, cataracts,weakened bones and skin, high blood pressure, elevated blood sugar, easybruising and decreased growth in children. Such side effects indicate aneed to accurately assess the efficacy of corticosteroid treatment inasthmatic patients.

Bronchodilators, also called “β2-agonists”, are non-steroidalanti-inflammatory medications often used as short-term “rescue”medications to immediately relieve asthma symptoms. Bronchodilatorsinclude albuterol, bitolterol, pirbuterol and terbutaline. Additionally,salmeterol is a long-acting β2-agonist that is intended to be used on along-term basis, along with an anti-inflammatory medication, forcontrolling asthma. Those using salmeterol should take the medication ona daily basis, even if they are feeling fine, as it prevents symptoms.Although sporadically effective, bronchodilators are not typicallyuseful in cases of severe asthma.

Many of the cells involved in causing airway inflammation are known toproduce signaling molecules within the body called “leukotrienes.”Leukotrienes are responsible for causing the contraction of the airwaysmooth muscle, increasing leakage of fluid from blood vessels in thelung, and further promoting inflammation by attracting otherinflammatory cells into the airways. Oral anti-leukotriene medicationshave been introduced to fight the inflammatory response typical ofallergic disease. These drugs are used in the treatment of chronicasthma. Recent data demonstrates that prescribed anti-leukotrienemedications can be beneficial for many patients with asthma, however, asignificant number of patients do not respond to anti-leukotriene drugs.

The present invention relates, in part, to methods for inducing apro-inflammatory like state in cultured cells. In a preferredembodiment, this involves increasing the level of particular pro-asthmalike effectors (e.g., cytokines such as, for example, IL-1β and TNFα).Pro-asthma like characteristics include, for example, heightenedconstrictor responsiveness to cholinergic stimulation and impairedrelaxation to β-adrenergic receptor stimulation ans airway inflammation.Accordingly, airway smooth muscle (“ASM”) contractility in the presenceof acetylcholine (“ACh”), and impaired ASM relaxation in response toisoproterenol, for example, can be taken as examples of pro-asthma and,thereby, pro-inflammatory like characteristics. Characteristics furtherinclude those that are exhibited in asthmatic patients or patients withan inflammatory disease. Such characteristics can be used to determinethe pro-asthma/pro-inflammatory like state. An assay for a subset or allknown characteristics can be used to determine apro-asthma/pro-inflammatory like state in resident tissue samples andcultured cells.

While infiltration of the airways with inflammatory cells, principallyinvolving eosinophils, mast cells, and lymphocytes, is implicated in theetiology of the altered airway responsiveness, recent studies havedetermined that, under specific conditions, the airway smooth muscle(ASM) itself has the capacity to autologously induce changes in itsconstrictor and relaxant responsiveness secondary to the induced releaseand autocrine actions of certain pro-inflammatory cytokines. Comparableautologous mechanisms have also been shown to exist in variousepithelial cells, keratinocytes, synovial cells, glial cells and villousintestinal cells. The present invention relates, in part, to methods forinducing a pro-inflammatory (e.g., pro-asthma like state) like state inresident tissue samples and cultured cells. Such cells can be, forexample ASM cells, epithelial cells, keratinocytes, synovial cells,glial cells and villous intestinal cells.

A pro-asthma like phenotype is associated with, for example, elevatedlevels of at least one effector such as certain cytokines, includingtumor necrosis factor alpha (hereinafter, “TNFα”) and interleukin 1-beta(hereinafter, “IL-1β”). These cytokines are also elevated patients withother inflammatory diseases, and, thus, cells induced to mimic apro-asthma/pro-inflammatory like state can be used to model, ex vivo,inflammatory disease generally (including, but not limited to, asthma,atopy (e.g., rhinitis, conjunctivitis, dermatitis, eczema), rheumatoidarthritis, psoriasis, IBD and COPD). As other effectors (e.g.,cytokines, signaling molecules, chemical and physical stimuli) arecharacterized, the methods described herein can be used in conjunctionwith additional effectors to induce the pro-asthma like state. In apreferred embodiment, the invention relates, in part, to methods forelevating the cellular level of TNFα and IL-1β, thereby inducingasthma-like or comparable changes in the responsiveness of the cells.Such cells, since they can be taken as a model for inflammatory disease,can be used in screening for drugs, screening for informative genes, andobtaining expression profiles for genes indicative of asthma and otherinflammatory diseases. It should be understood that any cytokine thatinduces a pro-inflammatory like state by exerting an inductive effect onTNFα and IL-1β expression and activity can be used as alternatives to orin combination with TNFα and/or IL-1β.

The cellular levels of TNFα and IL-1β can be increased by a variety ofmethods known in the art. For example, mammalian cells, such as ASMcells, epithelial cells, keratinocytes, synovial cells, glial cells andvillous intestinal cells grown in culture can be exposed to isolated andpurified TNFα and IL-1β such that these cytokines are taken up by thecells (typically, exposure of about 4 hours of TNFα at a concentrationof 5 ng/mL and IL-1β at a concentration of 1 ng/mL in culture willproduce pro-asthma/pro-inflammatory like symptoms in cultured cells.Other methods for expression of cytokines in cells grown in culture,e.g., by transfection of genes cloned into expression vectors, or bycontacting cells with effectors that are known to induce particularcytokines, are known in the art, and would allow for a similar inductionof pro-asthma/pro-inflammatory or pro-inflammatory like conditions incells.

The present invention also relates to methods for determining genes thatregulate responses to anti-inflammatory drugs, e.g., corticosteroids,used to treat inflammatory diseases such as asthma. The particulargenes, herein referred to as “informative genes,” are identified incells that have been induced to mimic asthma or other inflammatoryconditions. A subset or all informative genes can be assayed for geneexpression in order to generate an “expression profile” that includesgenes that regulate drug responses. As used herein, an “expressionprofile” refers to the level or amount of gene expression of one or moreinformative genes in a given sample of cells at one or more time points.A “reference” expression profile is a profile of a particular set ofinformative genes under particular conditions such that the expressionprofile is characteristic of a particular condition. As used herein,“gene” is a term used to describe a genetic element that gives rise toexpression products (e.g., pre-mRNA, mRNA, and polypeptides). Forexample, a reference expression profile that quantitatively describesthe expression of the informative genes listed in Tables 1 and 2 can beused as a reference expression profile for drug treatment and used toidentify genes that regulate drug responses. Thus by comparing geneexpression from a cell or tissue samples exposed to certainanti-inflammatory drugs with these reference expression profiles, genesthat regulate drug responses can be identified.

The generation of an expression profile requires both a method forquantitating the expression from informative genes and a determinationof the informative genes to be screened. The present invention describesscreening changes in individuals that affect the expression levels ofgene products in cells. As used herein, “gene products” aretranscription or translation products that are derived from a specificgene locus. The “gene locus” includes coding sequences as well asregulatory, flanking and intron sequences. Expression profiles aredescriptive of the level of gene products that result from informativegenes present in cells. Methods are currently available to one of skillin the art to quickly determine the expression level of several geneproducts from a sample of cells. For example, short oligonucleotidescomplementary to mRNA products of several thousand genes can bechemically attached to a solid support, e.g., a “gene chip,” to create a“microarray.” Specific examples of gene chips include Hu95GeneFL(Affymetrix, Santa Clara, Calif.) and the 6800 human DNA gene chip(Affymetrix, Santa Clara, Calif.). Such microarrays can be used todetermine the relative amount of mRNA molecules that can hybridize tothe microarrays (Affymetrix, Santa Clara, Calif.). This hybridizationassay allows for a rapid determination of gene expression in a cellsample. Alternatively, methods are known to one of skill in the art fora variety of immunoassays to detect protein gene expression products.Such methods can rely, for example, on conjugated antibodies specificfor gene products of particular informative genes. Described herein aremethods for identifying genes that regulate responses to inflammatorydrugs used to treat inflammatory diseases. In an attempt to furtherelucidate those genes that may contribute to pro-asthmatic changes inASM responsiveness, the effects of glucocorticoid treatment on ASMresponsiveness and its. associated pattern of altered gene expression inASM cells exposed to IL-1β and TNFα can be examined. Glucocorticoidsameliorate asthma symptoms and altered responsiveness in asthmaticairways, as well as the ability of glucocorticoids to generallyattenuate the expression of pro-inflammatory genes.

The identification of informative genes can be performed or verifiedunder ex vivo conditions. For example, pro-inflammatory like conditionscan be inducibly established in cultured cells. Described herein aremethods, for example, for producing cells with pro-inflammatory likecharacteristics. Such cells can be used to obtain reference expressionprofiles. In one embodiment of the present invention, cells induced toexhibit pro-inflammatory like characteristics are used to first identifyinformative genes that exhibit altered gene expression in response toanti-inflammatory drugs in diseases such as, for example, asthma.Expression profiles can be obtained for induced cells that have beenexposed to particular therapeutic agents (e.g., glucocorticoids such asdexamethasone or methylprednisolon), thus enabling identification ofgenes that are involved in regulating drug response.

IL-1β- and TNFα-related pathologies or diseases, as would be mimicked bythe pro-inflammatory like state induced in the cells described herein,include, but are not limited to, inflammatory diseases or disorders,infections, neurodegenerative diseases, malignant pathologies, cachecticsyndromes and certain forms of hepatitis.

Inflammatory diseases or disorders, include, but are not limited to,acute and chronic immune and autoimmune pathologies, such as, but notlimited to, rheumatoid arthritis (RA), juvenile chronic arthritis (JCA),psoriasis, graft versus host disease (GVHD), scleroderrna, diabetesmellitus, allergy; asthma, acute or chronic immune disease associatedwith an allogenic transplantation, such as, but not limited to, renaltransplantation, cardiac transplantation, bone marrow transplantation,liver transplantation, pancreatic transplantation, small intestinetransplantation, lung transplantation and skin transplantation; chronicinflammatory pathologies such as, but not limited to, sarcoidosis,chronic inflammatory bowel disease, ulcerative colitis, and Crohn'spathology or disease; vascular inflammatory pathologies, such as, butnot limited to, disseminated intravascular coagulation, atherosclerosis,Kawasaki's pathology and vasculitis syndromes, such as, but not limitedto, polyarteritis nodosa, Wegener's granulomatosis, Henoch-Schonleinpurpura, giant cell arthritis and microscopic vasculitis of the kidneys;chronic active hepatitis; Sjögren's syndrome; psoriatic arthritis;enteropathic arthritis; reactive arthritis and arthritis associated withinflammatory bowel disease; and uveitis.

Infections include, but are not limited to, sepsis syndrome, cachexia(e.g., TNFα-mediated effects), circulatory collapse and shock resultingfrom acute or chronic bacterial infection, acute and chronic parasiticand/or infectious diseases, bacterial, viral or fungal, such as a humanimmunodeficiency virus (HIV), acquired immunodeficiency syndrome (AIDS)(including symptoms of cachexia, autoimmune disorders, AIDS dementiacomplex and infections).

Neurodegenerative diseases include, but are not limited to,demyelinating diseases, such as multiple sclerosis and acute transversemyelitis.

Malignant pathologies are associated with TNFα-secreting tumors or othermalignancies involving TNFα, such as, for example, leukemias (acute,chronic myelocytic, chronic lymphocytic and/or myelodyspastic syndrome)and lymphomas (Hodgkin's and non-Hodgkin's lymphomas, such as malignantlymphomas (Burkitt's lymphoma or Mycosis fungoides)).

Cachectic syndromes and other pathologies and diseases involving excessTNFα, include, but not limited to, cachexia of cancer, parasitic diseaseand heart failure.

Elevated levels of TNFα are also associated with certain types ofhepatitis, including, but not limited to, alcohol-induced hepatitis andother forms of chronic hepatitis.

One of skill in the art will recognize that reagents necessary toutilize certain methods described herein can be contained in a kit. Suchreagents as described are either commercially available (e.g., bufferedsolutions, chemical reagents) or produced by methods known in the art(e.g., oligonucleotides, antibodies, ligands for detection). Thus, oneof skill in the art would recognize that a kit can be producedcontaining in appropriate compartments, for example, all reagents,probes, and materials necessary for to allow for the practice of themethods described herein.

The invention will be further described with reference to the followingnon-limiting examples. The teachings of all the patents, patentapplications and all other publications and websites cited herein areincorporated by reference in their entirety.

EXEMPLIFICATION Example 1 Ex vivo Model for Pro-Inflammatory Like State

Elevated levels of the pleiotropic cytokines, IL-1β and TNFα, have beenimplicated in the pathophysiology of asthma and other inflammatorydisorders (Broide, D. et al., 1992. J. Allergy Clin. Immunol.89:958-967; Arend, W., 2001. Arthritis Rheum. 45:101-106). To elucidatethe role of the cytokines IL-1β and TNFα in contributing to thepro-asthma like state, the effects of these cytokines on airway smoothmuscle (ASM) responsiveness and ASM multiple gene expression, assessedby high-density oligonucleotide array analysis, were examined in theabsence and presence of the glucocorticoid, dexamethasone (DEX).

In brief, administration of IL-1β/TNFα elicited increased ASMcontractility to acetylcholine (ACh) and impaired ASM relaxation toisoproterenol. These pro-asthmatic like changes in ASM responsivenesswere associated with IL-1β/TNFα induced upregulated mRNA expression of ahost of pro-inflammatory genes that regulate gene transcription,cytokines and chemokines, cellular adhesion molecules, and varioussignal transduction molecules that regulate ASM responsiveness. In thepresence of DEX, the induced changes in ASM responsiveness wereabrogated, and most of the IL-1β/TNFα mediated changes inpro-inflammatory gene expression were repressed (Table 1), although mRNAexpression of a small number of genes was further enhanced by DEX (Table2). Collectively, the observations supports the novel concept that,together with its role as a regulator of airway tone, in response toIL-1β/TNFα, ASM expresses a host of glucocorticoid-sensitive genes thatcontribute to the altered structure and function of airways in thepro-asthmatic state. The glucocorticoid-sensitive, cytokine-induced genepathways involved in ASM cell signaling represent important potentialtargets for new therapeutic interventions.

Animals

Ten adult New Zealand White rabbits were used in this study which wasapproved by the Biosafety and Animal Research Committee of the JosephStokes Research Institute at Children's Hospital of Philadelphia. Theanimals had no signs of respiratory disease prior to this study.

Preparation of ASM tissues

After anesthesia with xylazine (10 mg/kg) and ketamine (50mg/kg), theanimals were sacrificed with systemic air embolism. The tracheas wereremoved via open thoracotomy, cleared of loose connective tissue,divided into eight ring segments of 6-8 mm in length, and incubated for18 hr at room temperature in Dulbecco's modified Eagle's mediumcontaining both IL-1β (10 ng/mL) and TNFα (100 ng/mL), or in mediumalone with no added cytokines, both conditions in the absence andpresence of dexamethazone (DEX; 10⁻⁵M). The medium was aerated with acontinuous supplemental O₂ mixture (95% O₂/5% CO₂) during the incubationphase.

Pharmacodynamic Studies

After incubation, each airway segment was suspended longitudinallybetween stainless steel triangular supports in siliconized 20-mL organbaths (Harvard Apparatus, Inc., South Natick, Mass.). The lower supportwas secured to the base of the organ bath, and the upper support wasattached via a gold chain to a force transducer (FT.03C; GrassInstrument Co., Quincy, Mass.) from which isometric tension wascontinuously displayed on a multichannel recorder. Care was taken toplace the membranous portion of the trachea between the supports tomaximize the recorded tension generated by the contracting trachealismuscle. The tissues were bathed in modified Krebs-Ringer solutioncontaining (in mM) 125 NaCl, 14 NaHCO₃, 4 KCl, 2.25 CaCl₂.2H₂O, 1.46MgSO₄.7H₂O, 1.2 NaH₂PO4.H₂O, and 11 glucose. The baths were aerated with5% CO₂ in oxygen (a pH of 7.35-7.40 was maintained, and the temperaturewas held at 37° C.). Passive resting tension of each tracheal smoothmuscle segment was set at 2.0 g after each tissue had been passivelystretched to a tension of 8 g to optimize the resting length of eachsegment. The tissues were allowed to equilibrate in the bath for 45minutes, at which time each tissue was primed with a 1 minute exposureto 10⁻⁴M acetylcholine (ACh). Cholinergic contractility was initiallyassessed in the ASM by cumulative administration of ACh in final bathconcentrations from 10⁻⁹ to 10⁻³M. Thereafter, following thoroughrinsing, each tissue segment was half-maximally contracted with ACh, andrelaxation dose-response relationships to cumulative administration ofisoproterenol (10⁻⁹ to 10⁻⁴M) were generated in paired IL-1β/TNFαtreated and control tissues in the absence and presence of co-treatmentwith DEX. The initial constrictor dose-response curves to ACh wereanalyzed in terms of the tissues' maximal isometric contractile force(T_(max)) and sensitivity to the agonist, expressed as the negativelogarithm of the concentration of ACh producing 50% of T_(max) (pD₅₀;i.e., geometric mean ED₅₀ value). The relaxant responses toisoproterenol were analyzed in terms of percent maximal relaxation(R_(max)) from the active cholinergic contraction, and sensitivity tothe relaxing agent was determined as the corresponding pD₅₀ valueassociated with 50% R_(max).

Description of Microarray Gene Expression Studies

Simultaneous multiple gene mRNA expression was examined in human ASMcells with the Affymetrix expression microarray system using human genechips (HU95GeneFL array; Affymetrix, Santa Clara, Calif.) representingapproximately 6000 genes. The ASM cells were derived from a 21 year oldmale donor (Clonetics Corp., San Diego, Calif.) who had no evidence ofpulmonary disease, and the cells were carefully characterized by themanufacturer with specific markers to confirm their selective smoothmuscle phenotype and to exclude contamination with other cell types. Thecells were maintained at 37° C. in a humidified atmosphere of 5% CO₂/95%air and grown in a mixture of 5% Smooth muscle Basal Medium (SmBM),which was supplemented with 10% FBS, insulin (5 ng/mL), EGF (10 ng/mL;human recombinant), FGF (2 ng/mL; human recombinant), gentamycin (50ng/mL), and amphotericin-B (50 ng/mL). Once the cells reached -95%confluency, they were exposed for 4 hours to IL-1β (1 ng/mL) and TNFα (5ng/mL) combined, or to media alone in the absence and presence of 1 hourpre-treatment with DEX (10⁻⁵M).

Following incubation of the cells, the total RNA used for the Affymetrixmicroarray expression analysis was extracted and purified usingcommercially available reagents and in accordance with methodsrecommended by the manufacturer (Affymetrix, Santa Clara, Calif.).Briefly, total RNA was extracted using Trizol and purified with QiagenRNAEASY spin columns (Qiagen GmbH, Germany). Approximately 5 μg of RNAwas used for first and second strand cDNA synthesis. Afterprecipitation, the cDNAs were transcribed to cRNAs. The biotinylatedcRNA was subsequently hybridized to the Affymetrix gene chips overnight.Non-bound probes were removed by stringency washing. The hybridizedchips were developed using a Streptavidin-Phycoerythrin complex andscanned. The scanned images were then analyzed with Affymetrix softwareand the data was examined using commercially available softwareprograms, including Spotfire Net 5.1 (Spotfire Inc, Mass.).

Description of the Effects of IL-1β and TNFα on ASM Responsiveness

ASM constrictor dose-response relationships to ACh were determined inASM tissues pre-incubated for 24 hours in medium alone and in thepresence of maximally effective concentrations of IL-1β and TNFα. Asshown in FIG. 1, relative to controls, the IL-1β/TNFα treated tissuesexhibited significantly increased constrictor responsiveness to ACh,with mean±SE values for maximal isometric force of contraction (T_(max))amounting to 119.4±14.5 g/g ASM weight in the IL-1β/TNFα treated ASM but93.7±8.9 in the control ASM (p<0.01). Additionally, constrictorsensitivity to ACh was also relatively enhanced in the cytokine-treatedtissues, with mean±SE values for pD₅₀ (i.e.,−log ED₅₀) amounting to4.95±0.06−log M in the IL-1β/TNFα treated ASM but 4.66±0.12 in thecontrol ASM (p<0.05).

In separate studies, during comparable levels of initial sustainedACh-induced contractions, averaging ˜50% of T_(max), ASM relaxationresponses to cumulative administration of the beta-adrenergic agonist,isoproterenol, were generated in control and IL-1β/TNFα treated tissues.As shown in FIG. 2, relative to controls, the maximal relaxation(R_(max)) responses and pD₅₀ values for isoproterenol were significantlyattenuated in the IL-1β/TNFα treated tissues. Accordingly, the R_(max)values amounted to 41.3±6.0 in the cytokine-treated ASM and 57.7±7.1% inthe control ASM (p<0.01), and the corresponding pD₅₀ values amounted to5.87±0.05 and 6.09±0.11−log M, respectively (p<0.05).

Description of the Effects of IL-1β/TNFα on ASM Cell Gene Expression

In light of the above observations, to elucidate potential gene pathwaysassociated with IL-1β/TNFα induced changes in ASM responsiveness, theeffects of these cytokines on mRNA expression of multiple genesputatively involved in various cell signaling processes in ASM wereexamined in cultured human ASM cells. Accordingly, using a high densityoligonucleotide DNA microarray analysis, in 4 separate experiments, itwas determined that ˜40% of genes were expressed in untreated ASM cells,and that treatment of cells with IL-1β/TNFα did not significantly alterthe total number of genes expressed. More than 400 genes, however,demonstrated up- or down-regulation of their mRNA signals in response toIL-1β/TNFα administration. Given the established sensitivity of theexpression technique applied, a two-fold increase in signal intensitiesfrom baseline was considered significant. Accordingly, ˜70 genes thatplay a potential role in cell signaling in ASM demonstrated 22-fold(i.e., 2 to ˜150-fold) increase in mRNA expression in response toIL-1β/TNFα. The latter collection of genes is categorically displayed inFIGS. 3-6, with the genes in each category identified by their symbolsand GenBank accession number, and plotted in relation to theirrespective magnitudes (mean±SE values of fold-increase) of altered mRNAexpression.

Within the cytokine/chemokine category of genes, those depictingupregulated mRNA expression by an average of ≧10-fold above baseline inresponse to IL-1β/TNFα included the small inducible cytokine sub-familyB (SCYB) members -2, -3, -1, -5, and -6, IL-1β, IL-8, CSF-2 (i.e.,GM-CSF), TNFα-IP3, IL-6, and CSF-3 (FIG. 3). Within the cellularadhesion molecule (CAM)/extracellular matrix (ECM)-related category ofgenes, those upregulated ≧10-fold included ICAM-1, matrixmetalloproteinase (MMP)-12, and VCAM-1 (FIG. 4), and, within thecategory of transcription factors, the genes comparably upregulatedincluded NR4A3 and BCL-2A1 (FIG. 5). Other genes related to variousaspects of cellular signaling/metabolism, including those encodingvarious proteases, kinases, and other molecules involved in signaltransduction, were also upregulated in ASM cells in response IL-1β/TNFαadministration (FIG. 6), most notably including phosphodiesterase 4B(PDE-4B), superoxide dismutase 2 (SOD-2), and inducible cyclooxygenase 2(COX-2). Contrasting these observations, treatment of cells withIL-1β/TNFα had no effect on mRNA expression of constitutively expressed“house-keeping” genes such as β-actin, ribosomal protein L7,β2-microglobulin, and others.

Description of the Effects of Glucocorticoids on IL-1,/TNFα InducedChanges in ASM Responsiveness

To assess whether the IL-1β/TNFα induced changes in ASM responsivenessare glucocorticoid-sensitive, contractile dose-response relationships toACh were compared between IL-1β/TNFα treated ASM tissues and theirrespective paired control ASM segments, both in the absence and presenceof pretreatment of the tissues for 1 hour with dexamethasone (DEX;10⁻⁵M). As shown in FIG. 7, the heightened constrictor responses to AChgenerated in IL-1β/TNFα exposed ASM were abrogated by pre-treating thecytokine-exposed tissues with DEX. Accordingly, in these DEX-pre-treatedtissues, the mean±SE T_(max) and pD₅₀ values amounted to 102.9±13.1 g/gASM weight and 4.89±0.05−log M, respectively, and the latterdeterminations were similar to those obtained in control ASM. Incontrast, pretreatment with DEX had no effect on the constrictorresponses to ACh in control tissues (FIG. 7; open squares).

Comparable to the above protective effects of DEX on cytokine-inducedchanges in ASM constrictor responsiveness, the impairedbeta-adrenoceptor-mediated relaxation responses to isoproterenolobtained in IL-1β/TNFα exposed ASM were also completely abrogated bypre-treating the tissues with DEX (FIG. 8). Accordingly, in theseDEX-pre-treated tissues, the mean R_(max) and pD₅₀ values forisoproterenol averaged 55.5±5.7% and 5.99±0.06−log M, respectively; andthe latter determinations were similar to those obtained in control ASM.In contrast, pretreatment with DEX had no effect on the relaxationresponses to isoproterenol in control tissues (FIG. 8; open squares).

Description of the Effects of Glucocorticoids on IL-1β/TNFα Induced GeneExpression in ASM Cells

Given the efficacy of DEX in ablating the effects of IL-1β/TNFα on ASMresponsiveness, the ability of DEX to modulate the above observedeffects of IL-1β/TNFα on multiple gene expression in ASM cells wasexamined. Paired cultures of ASM cells were exposed to media alone(control) or to IL-1β/TNFα in the absence and presence of DEX (10⁻⁵M),with each condition examined in duplicate. For any given gene,sensitivity to DEX was then determined as the ratio of the altered mRNAlevels elicited by IL-1β/TNFα in the presence/absence of DEX.Accordingly, a mRNA expression ratio (MER) of 1.0 implies a lack ofeffect of DEX, whereas MER values below and above 1.0 denote DEX-inducedrepression and stimulation of mRNA expression, respectively. The resultsdemonstrate that the upregulated mRNA levels exhibited by cells exposedto IL-1β/TNFα were largely repressed by pre-treating the cells with DEX,as evidenced by MER values below 1.0 for the majority of genes belongingto each category (FIG. 9). Not all genes, however, displayed DEXsensitivity and, as further shown in FIG. 9, a small number of genes ineach category exhibited stimulation of IL-1β/TNFα induced mRNAexpression in the presence of DEX (i.e., MER>1.0). In evaluating thevariability in DEX sensitivity within each category of genes, nocorrelation was found between MER values of the different genes and thecorresponding magnitudes of IL-1β/TNFα induced enhanced mRNA expressionin the absence of DEX.

The DEX-sensitive genes depicted in FIG. 9 that exhibit ≧10% DEX-induceddecrease in mRNA expression (i.e., MER ≦0.90) are identified in Table 1.It will be noted that a strong repressive effect of DEX was seen forgenes known to be involved in the regulation of cAMP and Ca²⁺mobilization, including the phosphodiesterase D4 and plasma membraneCa²⁺ ATPase genes, which provided MER values of 0.40 and 0.34,respectively, corresponding to 60% and 66% inhibition of IL-1β/TNFαinduced mRNA expression in the presence of DEX, respectively.Additionally, certain cytokine/chemokine-related and other cellsignaling-related genes were significantly inhibited by DEX, includingthe pro-IL-1β, IL-8, IL-13R, small inducible cytokine subfamily(SCY)-B2, -B6, -A7, and bradykinin receptor-2 (BKR2) genes. Moreover, itis relevant to note that the p50-NF-kB gene, which belongs to the NF-kBfamily of inducible transcription factors that regulates the host immuneand inflammatory responses, was inhibited by 51% by DEX (i.e.,MER=0.49). Finally, MMP3, MMp10 and MMp12, genes which are importantlyimplicated in tissue remodeling, were also markedly inhibited by DEX.TABLE 1 Genes repressed by DEX in IL-1β/TNFα treated ASM cells Name/Gene Category GenBank MER CAM/ECM Molecules Intercellular adhesionmolecule-1 ICAM-1/M24283 0.66 Matrix metalloproteinase-3 MMP3/X052320.70 Matrix metalloproteinase-10 MMP10/X07820 0.00 Matrixmetalloproteinase-12 MMP12/L23808 0.20 Ninjurin-1 NINJ1/U72661 0.59 CD83CD83/Z11697 0.56 Cytokines/Chemokines pro-Interleukin-1β IL-1β/X045000.45 Small inducible cytokine subfamily B3 SCYB3/X53800 0.83 ColonyStimulating factor-2 CSF2/M13207 0.53 RANTES RANTES/M21121 0.67Interleukin-6 IL-6/X04602 0.89 Fibroblast growth factor-2 FGF2/J045130.69 Interleukin-8 IL-8/M28130 0.57 Small inducible cytokine subfamilyB2 SCYB2/M57731 0.52 Inhibin INHBA/X57579 0.00 Interleukin-13 receptor-αIL-1 RA2/U70981 0.00 Small inducible cytokine subfamily B6 SCYB6/U833030.61 Small inducible cytokine subfamily A7 SCYA7/X72308 0.50Monocyte-derived chemotactic protein MDCP/HG4069 0.29 Pre-B cellcolony-enhancing factor PBEF/U02020 0.73 Cell Signaling/MetabolismPhosphodiesterase-4B PDE4B/L20971 0.40 Adenosine monophosphate deaminaseAMPD3/D12775 0.82 Urokinase plasminogen activator uPA/X02419 0.00Bradykinin receptor-2 BKR2/X86163 0.62 CDC28 protein kinase-2CKS2/X54942 0.80 Plasma membrane Ca²⁺ ATPase ATP2B1/J04027 0.34Superoxide dismutase-2 SOD2/X07834 0.89 Transcription Factors Nuclearfactor of kappa light polypeptide NFKB1/M58603 0.87 Nuclear factor ofactivated T cells-C1 NFATC1/U08015 0.61 BCL2-related protein A1BCL2A1/U29680 0.41 Signal transducer/activator of transcription 5STAT5A/U43185 0.87 Nef-associated factor 1 NAF1/D30755 0.88 Nuclearreceptor subfamily 4 NR4A3/U12767 0.54 Putative lymphocyte G0/G1 switchgene G0S2/M72885 0.65 p50-NF-KB p50-NF-kB/S76638 0.49 PTX3 promoterPTX3/X97748 0.69Table 1. Genes repressed by dexamethasone (DEX) in IL-1β/TNFα treatedhuman ASM cells. Genes are identified by their gene symbols and GenBankaccession numbers. The MER value given for each gene refers to the ratioof mRNA expression levels elicited by IL-1β/TNFα in the presence versusabsence of DEX.

Among the collection of DEX-sensitive genes exhibiting MER values≧1.10,as shown in Table 2, those belonging the cell signaling-related categoryincluded 11-beta-hydroxysteroid-dehydroxygenase-1, the MAP kinasesubtype, MAPKKK5, and the ATP-binding cassette gene, ABC-B2. In thecytokine/chemokine-related category, DEX-induced augmented mRNAexpression was most evidenced by genes encoding epithelial-derivedneutrophil-activating peptide 78 (SCYB5), colony (granulocyte)stimulating factor 3 (CSF3), TNFα induced protein 3 (TNFα-IP3) andTNFα-IP6.

Other genes upregulated by DEX include the transcription factor-relatedgene, CCAAT-enhancer binding protein (C-EBP)-delta, and the CAM/ECMmolecule-related gene, tenascin C. TABLE 2 Genes stimulated by DEX inIL-1β/TNFα treated ASM cells Name/ Gene Category GenBank MER CAM/ECMMolecules Tenascin C HXB/X78565 1.21 Cytokines/Chemokines Smallinducible subfamily B5 SCYB5/L37036 1.75 Tumor necrosis factor-α-inducedprotein 6 TNF-αIP6/M31165 1.11 Tumor necrosis factor-α-induced protein 3TNF-αIP3/M59465 1.21 Small inducible cytokine subfamily B10SCYB10/X02530 1.12 Colony stimulating factor-3 CSF-3/X03656 1.31 CellSignaling/Metabolism Guanylate binding protein-1 GBP1/M55542 1.22Hydroxysteroid (11-β) dehydrogenase-1 HSD11B1/M7665 2.57 ATP-bindingcassette ABCB2/X57522 1.75 Mitogen-activated protein kinase kinaseMAP3K5/U67156 1.65 kinase-5 Transcription Factors KIAA0247 gene productKIAA0247/D87434 1.3 Enhancer binding protein CEBPD/M83667 1.48Table 2. Genes stimulated by DEX in IL-1β/TNFα treated ASM cells. Genesare identified by their gene symbol and GenBank accession numbers. TheMER value given for each gene refers to the ratio of mRNA expressionlevels elicited by IL-1β/TNFα in the presence or absence of DEX.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

1. A method for inducing a pro-inflammatory like state in a residenttissue cell, comprising contacting the cell with one or more cytokinesthat induce a pro-inflammatory like state.
 2. The method of claim 1,wherein the cytokine is IL-1β, TNFα or both.
 3. The method of claim 1,wherein the resident tissue cell is selected from the group consistingof: airway smooth muscle cell, epithelial cell, keratinocyte, synovialcell, glial cell and villous intestinal cell.
 4. The method of claim 1,wherein the pro-inflammatory like state is a pro-asthma like state. 5.The method of claim 4, wherein the resident tissue cell is airway smoothmuscle or airway epithelial cell.
 6. A resident tissue cell induced toexhibit a pro-asthma like state according to the method of claim
 4. 7.The resident tissue cell of claim 6, wherein the resident tissue cell isairway smooth muscle or airway epithelial cell.
 8. A method forscreening drug candidates for treating an inflammatory disease,comprising: a) contacting a resident cell induced by the method of claim1 with a drug candidate for treating the inflammatory disease; and b)assaying for a pro-inflammatory like state, such that an absence of thepro-inflammatory like state is indicative of the drug candidate beingeffective in treating the inflammatory disease.
 9. The method of claim8, wherein the inflammatory disease is selected from the groupconsisting of: asthma, atopy, rheumatoid arthritis, psoriasis,inflammatory bowel disease and chronic obstructive pulmonary disease.10. The method of claim 9, wherein the atopy is selected from the groupconsisting of rhinitis, conjunctivitis, dermatitis and eczema.
 11. Amethod for inducing a resident tissue cells to mimic an inflammatorydisease, comprising increasing expression of IL-1β, TNFα or both in thecells.
 12. The method of claim 11, wherein the inflammatory disease isasthma.
 13. The cell of claim 12, wherein the resident tissue cell isairway smooth muscle or airway epithelial cell.
 14. A resident tissuecell induced according to the method of claim
 11. 15. The method ofclaim 11, wherein the resident tissue cell is selected from the groupconsisting of: airway smooth muscle cell, epithelial cell, keratinocyte,synovial cell, glial cell and villous intestinal cell.
 16. A method forscreening drug candidates for treating an inflammatory disease,comprising: a) contacting the resident tissue cell induced according tothe method of claim 11 with a drug candidate for treating aninflammatory disease; and b) assaying for a pro-inflammatory like state,such that an absence of the pro-inflammatory like state is indicative ofthe drug candidate being effective in treating an inflammatory disease.17. The method of claim 16, wherein the inflammatory disease is selectedfrom the group consisting of: asthma, atopy, rheumatoid arthritis,psoriasis, inflammatory bowel disease and chronic obstructive pulmonarydisease.
 18. The method of claim 17, wherein the atopy is selected fromthe group consisting of rhinitis, conjunctivitis, dermatitis and eczema.19. A method of identifying genes associated with an inflammatorydisease, comprising: a) obtaining resident tissue cells induced to mimicthe inflammatory disease; b) assaying the expression level of at leastone gene in the cells; c) comparing the expression level from b) to thebaseline expression levels in cells not induced to mimic theinflammatory disease; and d) identifying a difference in expressionlevel in cells induced to mimic the inflammatory disease versus cellsthat do not mimic the inflammatory disease, such difference indicatingthe gene of b) is associated with the inflammatory disease.
 20. A methodfor identifying a gene that regulates drug response in inflammatorydisease, comprising: a) obtaining a gene expression profile for at leastone informative gene identified by the method of claim 19 in a residenttissue cell induced for a pro-inflammatory like state in the presence ofthe candidate drug; and b) comparing the expression profile of theinformative gene to a reference expression profile for the informativegene in a cell induced for the pro-inflammatory like state in theabsence of the candidate drug, wherein genes whose expression relativeto the reference expression profile is altered by the drug mayidentifies the gene as a gene that regulates drug response ininflammatory disease.
 21. An informative gene identified by the methodof claim
 19. 22. The method of claim 21, wherein the informative gene isselected from the genes described in Tables 1 and
 2. 23. A method fordiagnosing an inflammatory disease, comprising: a) obtaining a geneexpression profile for a resident tissue cell induced to mimic apro-inflammatory like state for at least one informative gene identifiedby the method of claim 20; b) comparing the expression profile of theinformative gene to a reference expression profile for the informativegene in a resident tissue cell induced for pro-inflammatory likeconditions in the presence of an anti-inflammatory drug, wherein thegenes that are induced and reversed by anti-inflammatory drug treatmentindicate the inflammatory disease.
 24. The method of claim 23, whereinthe informative gene is selected from the genes described in Tables 1and
 2. 25. An expression profile indicative of the presence of asthma ina patient, comprising at least one informative gene of Table 1 and Table2.